Substituted 10-aryl-11H-benzo[b]fluorenes and 7-aryl-5, 6-dihydro-benzo[a]anthracenes for selective effects on estrogen receptors

ABSTRACT

This invention provides a compound, or its possible salt, having the formula, 
                         
wherein: R e  and ′R e  are OH, optionally independently etherified or esterified; Z is —CH 2 CH 2 — or —C(R 4 ,R 5 )—, wherein R 4  and R 5  are independently H, (1C-2C)alkyl or form together a spiro(3C-5C)cycloalkyl; R 1  is H, halogen, CF 3 , or (1C-4C)alkyl; R 2  and R 3  are independently H, halogen, —CF 3 , —OCF 3 , (1C-8C)alkyl, hydroxy, (1C-8C)alkyloxy, aryloxy, aryl(1C-8C)alkyl, halo(1C-8C)alkyl, —O(CH 2 ) m X, wherein X is halogen or phenyl and m=2-4; —O(CH 2 ) m NR a R b , —S(CH 2 ) m NR a R b  or —(CH 2 ) m NR a R b , wherein m=2-4 and wherein R a , R b  are independently (1C-8C)alkyl, (2C-8C)alkenyl, (2C-8C)alkynyl, or aryl, which alkyl, alkenyl and aryl can optionally be substituted with halogen, —CF 3 , —OCF 3 , —CN, —NO 2 , —OH, (1C-8C)alkoxy, aryloxy, carboxyl, (1C-8C)alkylthio, carboxylate, (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl or halo(1C-8C)alkyl; or R a  and R b  form a 3-8 membered ring structure, optionally substituted with halogen, —CF 3 , —OCF 3 , —CN, —NO 2 , hydroxy, hydroxy(1C-4C)alkyl, (1C-8C)alkoxy, aryloxy, (1C-8C)alkylthio, carboxyl, carboxylate, (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl, halo(1C-8C)alkyl.

The invention relates to a non-steroidal compound with selectiveaffinity for estrogen receptors and to a method for selective estrogenreceptor modulation (SERM) with such a compound and to the use of such acompound for the manufacture of a medicine for estrogen-receptor relatedtreatments.

Compounds with affinity for estrogen receptors have found long-standingutility in the treatment of a variety of medical indications and inregimes for contraceptive purposes. Despite the long history of thefield there still is a need for more effective, safer and moreeconomical compounds than the existing ones. This need is the morepressing in view of advancement in health care in other areas, which hasled to an increasingly longer life span. This is in particular a problemfor women for whom the decline in estrogenic hormones at menopause isdrastic and has negative consequences for bone strength andcardiovascular functions. For the control or prevention of estrogensensitive tumor growth, compounds are needed which are antagonists,partial antagonists or tissue selective agonists for estrogen receptors.

The discovery of subtypes of estrogen receptors, there being anα-subtype (ERα) and a β-subtype (ERβ) of such receptors (Mosselman etal., FEBS Letters vol. 392 (1996) pp. 49-53 as well as EP-A-0 798 378),offers the possibility to influence one particular subtype of those tworeceptors more selectively, immanently resulting in more effectivetreatments or treatments with less side effects. Since these receptorshave a different distribution in human tissue, the finding of compoundswhich possess a selective affinity for either of the two is an importanttechnical progress, making it possible to provide a more selectivetreatment in estrogen-receptor related medical treatments, such as thosefor contraception and for treatment of menopausal complaints,osteoporosis, and estrogen dependent tumour control, with a lower burdenof estrogen-related side-effects.

In WO 01/72713 certain compounds with an unsaturated or partiallyunsaturated four-ring skeleton with hydroxyl substitutions at specificlocations, i.e. 2,8-dihydroxy-11H-benzo[b]fluorene and3,9-dihydroxy-5,6-dihydro-benz[a]anthracene are disclosed. Some arylsubstituted derivatives of these compounds possess high antagonism forERβ and may also show ERα antagonism or ERα agonism as is published inthe non-prepublished patent application WO 02/16316. In the latterdocument compounds are defined having the formula

wherein:

-   -   R^(e) and ′R^(e) are OH, optionally independently etherified or        esterified;    -   Y is —CH₂— or —CH₂CH₂—;    -   R¹ is H, halogen, CF₃, or (1C-4C)alkyl;    -   R⁶, R⁷ and R⁸ are independently H, halogen, —CF₃, —OCF₃,        (1C-8C)alkyl, hydroxy, (1C-8C)alkyloxy, aryloxy,        aryl(1C-8C)alkyl, halo(1C-8C)alkyl, —O(CH₂)_(m)X, wherein X is        halogen or phenyl and m=2-4; —O(CH₂)_(m)NR_(c)R_(d),        —S(CH₂)_(m)NR_(c)R_(d) or —(CH₂)_(m)NR_(c)R_(d), wherein m=2-4        and wherein R_(c), R_(d) are independently (1C-8C)alkyl,        (2C-8C)alkenyl, (2C-8C)alkynyl, or aryl, optionally substituted        with halogen, —CF₃, —OCF₃, —CN, —NO₂, —OH, (1C-8C)alkoxy,        aryloxy, carboxyl, (1C-8C)alkylthio, carboxylate, (1C-8C)alkyl,        aryl, aryl(1C-8C)alkyl, halo(1C-8C)alkyl or R_(c) and R_(d) form        a 3-8 membered ring structure, optionally substituted with        halogen, —CF₃, —OCF₃, —CN, —NO₂, hydroxy, hydroxy(1C-4C)alkyl,        (1C-8C)alkoxy, aryloxy, (1C-8C)alkylthio, carboxyl, carboxylate,        (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl, halo(1C-8C)alkyl.

This invention makes a compound available having the formula 1,

wherein:

-   -   R^(e) and ′R^(e) are OH, optionally independently etherified or        esterified;    -   Z is —CH₂CH₂— or —C(R⁴,R⁵)—, wherein R⁴ and R⁵ are independently        H, (1C-2C)alkyl or form together a spiro(3C-5C)cycloalkyl;    -   R¹ is H, halogen, CF₃, or (1C-4C)alkyl;    -   R² and R³ are independently H, halogen, —CF₃, —OCF₃,        (1C-8C)alkyl, hydroxy, (1C-8C)alkyloxy, aryloxy,        aryl(1C-8C)alkyl, halo(1C-8C)alkyl, —O(CH₂)_(m)X, wherein X is        halogen or phenyl and m=2-4; —O(CH₂)_(m)NR_(a)R_(b),        —S(CH₂)_(m)NR_(a)R_(b) or —(CH₂)_(m)NR_(a)R_(b), wherein m=2-4        and wherein R_(a), R_(b) are independently (1C-8C)alkyl,        (2C-8C)alkenyl, (2C-8C)alkynyl, or aryl, which alkyl, alkenyl        and aryl can optionally be substituted with halogen, —CF₃,        —OCF₃, —CN, —NO₂, —OH, (1C-8C)alkoxy, aryloxy, carboxyl,        (1C-8C)alkylthio, carboxylate, (1C-8C)alkyl, aryl,        aryl(1C-8C)alkyl or halo(1C-8C)alkyl; or R_(a) and R_(b) form a        3-8 membered ring structure, optionally substituted with        halogen, —CF₃, —OCF₃, —CN, —NO₂, hydroxy, hydroxy(1C-4C)alkyl,        (1C-8C)alkoxy, aryloxy, (1C-8C)alkylthio, carboxyl, carboxylate,        (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl, halo(1C-8C)alkyl.

It is thus found, as one embodiment of this invention, compounds havingthe formula 2

wherein:

-   -   R^(e) and ′R^(e) are OH, optionally independently etherifed or        esterified;    -   R¹ is H, halogen, CF₃, or (1C-4C)alkyl;    -   R² and R³ are independently H, halogen, —CF₃, —OCF₃,        (1C-8C)alkyl, hydroxy, (1C-8C)alkyloxy, aryloxy,        aryl(1C-8C)alkyl, halo(1C-8C)alkyl, —O(CH₂)_(m)X, wherein X is        halogen or phenyl and m=2-4; —O(CH₂)_(m)NR_(a)R_(b),        —S(CH₂)_(m)NR_(a)R_(b) or —(CH₂)_(m)NR_(a)R_(b), wherein m=2-4        and wherein R_(a), R_(b) are independently (1C-8C)alkyl,        (2C-8C)alkenyl, (2C-8C)alkynyl, or aryl, which alkyl, alkenyl        and aryl can optionally be substituted with halogen, —CF₃,        —OCF₃, —CN, —NO₂, —OH, (1C-8C)alkoxy, aryloxy, carboxyl,        (1C-8C)alkylthio, carboxylate, (1C-8C)alkyl, aryl,        aryl(1C-8C)alkyl or halo(1C-8C)alkyl; or R_(a) and R_(b) form a        3-8 membered ring structure, optionally substituted with        halogen, CF₃, —OCF₃, —CN, —NO₂, hydroxy, hydroxy(1C-4C)alkyl,        (1C-8C)alkoxy, aryloxy, (1C-8C)alkylthio, carboxyl, carboxylate,        (1C-8C)alkyl, aryl, axyl(1C-8C)alkyl, halo(1C-8C)alkyl;    -   R⁴ and R⁵ are independently (1C-2C)alkyl or form together a        spiro(3C-5C)cycloalkyl.

These compounds, possessing high selective antagonism for ERβ, can inaddition, have improved resistence to oxidative attack by having alkylsubstitution at position 11 of the 11H-benzo[b]fluorene skeleton.

The present invention further makes the following group of very usefulcompounds available, that is a compound having the formula 3

wherein:

-   -   R^(e) and ′R^(e) are OH, optionally independently etherified or        esterified;    -   Z is —CH₂CH₂— or —C(R³,R⁴)—, wherein R³ and R⁴ are independently        H, (1C-2C)alkyl or form together a spiro(3C-5C)cycloalkyl;    -   R¹ is H, halogen, CF₃, or (1C-4C)alkyl;    -   R² is —O(CH₂)_(m)NR_(a)R_(b), —S(CH₂)_(m)NR_(a)R_(b) or        —(CH₂)_(m)NR_(a)R_(b), wherein m=2-4 and wherein R_(a), R_(b)        are independently (1C-8C)alkyl, (2C-8C)alkenyl, (2C-8C)alkynyl,        or aryl, which alkyl, alkenyl and aryl can be optionally        substituted with halogen, —CF₃, —OCF₃, —CN, —NO₂, —OH,        (1C-8C)alkoxy, aryloxy, carboxyl, (1C-8C)alkylthio, carboxylate,        (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl or halo(1C-8C)alkyl; or        R_(a) and R_(b) form a 3-8 membered ring structure, optionally        substituted with halogen, —CF₃, —OCF₃, —CN, —NO₂, hydroxy,        hydroxy(1C-4C)alkyl, (1C-8C)alkoxy, aryloxy, (1C-8C)alkylthio,        carboxyl, carboxylate, (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl,        halo(1C-8C)alkyl.

More specific embodiments of the previously defined embodiments can beobtained by selecting H, halogen or CF₃ for R¹. Compounds according tothe formulas whereby R¹ is halogen, whereby chlorine is most preferred,are particularly potent and selective for the ERβ.

Another embodiment of the invention is a non-steroidal compound with a10-Aryl-11H-benzo[b]fluorene skeleton having the formula 4

wherein:

-   -   R^(e) and ′R^(e) are OH, optionally independently etherified or        esterified;    -   R¹ is H, halogen or CF₃;    -   R² is —O(CH₂)_(m)NR_(a)R_(b), —S(CH₂)_(m)NR_(a)R_(b) or        —(CH₂)_(m)NR_(a)R_(b), wherein m=2-4 and wherein R_(a), R_(b)        are independently (1C-8C)alkyl, (2C-8C)alkenyl, (2C-8C)alkynyl,        or aryl, which alkyl, alkenyl and aryl can be optionally        substituted with halogen, —CF₃, —OCF₃, —CN, —NO₂, —OH,        (1C-8C)alkoxy, aryloxy, carboxyl, (1C-8C)alkylthio, carboxylate,        (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl or halo(1C-8C)alkyl; or        R_(a) and R_(b) form a 3-8 membered ring structure, optionally        substituted with halogen, —CF₃, —OCF₃, —CN, —NO₂, hydroxy,        hydroxy(1C-4C)alkyl, (1C-8C)alkoxy, aryloxy, (1C-8C)alkylthio,        carboxyl, carboxylate, (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl,        halo(1C-8C)alkyl.

For compounds having formula 4 it is preferred to select those in which

-   -   R² is —O(CH₂)_(m)NR_(a)R_(b), wherein m=2-3 and R_(a), R_(b) are        independently (1C-5C)alkyl, (3C-5C)alkenyl, or aryl, which        alkyl, alkenyl and aryl can be optionally substituted with OH or        methoxy, or R_(a) and R_(b) form a 4-7 membered ring structure        selected from the list: azetidine, pyrrolidine, 3-pyrroline,        piperidine, piperazine, tetrahydropyridine, morpholine,        thiomorpholine, thiazolidine, homopiperdine, tetrahydroquinoline        and 6-azabicyclo[3.2.1]octane, which 4-7 membered ring structure        can optionally be substituted with OH, methoxy, acetyl,        carboxylate, (1C-3C)alkyl, phenyl, benzyl, and phenylethyl.

In those cases that a compound of the invention contains a basic aminefunction, the compound may be used as a free base or as apharmaceutically acceptable salt such as hydrochloride, acetate,oxalate, tartrate, citrate, phosphate, maleate or fumarate.

The ester and ether compounds in the collection of compounds accordingto the invention often have activity as prodrug. A prodrug is defined asbeing a compound which converts in the body of a recipient to a compoundas defined by the formulas 1 to 4 and to the free hydroxyl compounds ofthe above defined compounds. Preferred ester and ether prodrugs arecarboxylic acid esters or alkyl ethers on one or both hydroxyl groups,and more preferred prodrugs are (2C-6C)carboxylic acid esters, such asesters of (iso)butanoic acid, or (1C-4C) alkyl ethers. In general, thehydroxy groups can for example be substituted by allyl*oxy, alkenyl*oxy,acyl*oxy, aroyloxy, alk*oxycarbonyloxy, sulfonyl groups or phosphategroups, whereby the carbon chain length of the groups denoted with anasterisk (*) is not considered to be sharply delimited, while aroylgenerally will comprise a phenyl, pyridinyl or pyrimidyl, which groupscan have substitutions customary in the art, such as alkyl*oxy, hydroxy,halogen, nitro, cyano, and (mono-, or dialkyl*-)amino. The length of thealkyl, alkenyl and acyl groups is selected depending on the desiredproperties of the prodrugs, whereby the longer chained prodrugs with forexample lauryl or caproyl chains are more suitable for sustained releaseand depot preparations. It is known that such substituents spontaneouslyhydrolyse or are enzymatically hydrolysed to the free hydroxylsubstituents on the skeleton of the compound. Such prodrugs will havebiological activity comparable to the compounds to which they areconverted in the body of the recipients. The active compound to which aprodrug is converted is called the parent compound. The onset of actionand duration of action as well as the distribution in the body of aprodrug may differ from such properties of the parent compound.

Other terms used in this description have the following meaning:

-   alkyl is a branched or unbranched alkyl group, for example methyl,    ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, hexyl,    octyl, capryl, or lauryl;-   alkenyl is a branched or unbranched alkenyl group, such as ethenyl,    2-butenyl, etc.;-   alkynyl is a branched or unbranched alkynyl group, such as ethynyl    and propynyl;-   halogen refers to fluorine, chlorine, bromine and iodine;-   aryl is a mono- or polycyclic, homo- or heterocyclic aromatic ring    system, such as phenyl, naphtyl or pyridinyl; a monocyclic ring with    6 atoms is preferred for use;-   a ring system or structure is referring to a chemical group in which    all atoms are involved in formed rings, which rings can be saturated    or (partially) unsaturated and comprise C, O, S or N atoms;-   aroyl is arylcarbonyl such as a benzoyl group;-   alkanoyl means a formyl or alkylcarbonyl group such as formyl,    acetyl and propanoyl;-   acyl is a (substituent-)carbonyl group, such as an aroyl or    alkanoyl;-   carboxyl is a —COOH substituent, making the compound an organic    acid;-   carboxylate is an ester or salt of a carboxyl substituent;-   3-8 membered ring structure refers to a single ring or two or more    fused rings comprising 3-8 atoms, for example, when a nitrogen atom    is in the ring, an azetidine, pyrrolidine, 3-pyrroline, piperidine,    piperazine, tetrahydropyridine, morpholine, thiomorpholine,    thiazolidine, homopiperidine, tetrahydroquinoline or    6-azabicyclo[3.2.1]octane.

The prefixes (1C-4C), (2C-4C) etc. have the usual meaning to restrictthe meaning of the indicated group to those with 1 to 4, 2 to 4 etc.carbon atoms.

The estrogen-receptor affinity profile of the compounds according to thepresent invention, makes them suitable for use in estrogen-receptorrelated medical treatments, in the sense that these compounds areimproved selective anti-estrogens, partial anti-estrogen or partialestrogens. Estrogen-receptor related medical treatments specificallynamed are those for contraception or for treatment or prevention ofbenign prostate hypertrophy, cardiovascular disorders, menopausalcomplaints, osteoporosis, estrogen dependent tumour control or centralnervous system disorders such as depression or Alzheimer's disease. Inparticular the most selective compounds for the ERβ receptor aresuitable for estrogen-receptor related medical treatments underdiminished estrogen-related side-effects. This is most desirable whenthese compounds are used in the treatment of osteoporosis,cardiovascular disorders and central nervous system disorders such asdepression or Alzheimer's disease. Selective blockade of ERβ-receptorswith compounds of this invention can be used to prevent and reducemalignent tumor growth and hyperplasias. The receptor selectivity helpsto effectuate tissue selectivity. Those tissues rich in ERβ-receptorscan be protected by ERβ-receptor antagonists from the risk ofstimulation of growth by estrogenic agonists. The latter can be ofendogenous origin or from exogenous origine when administered duringestrogenic treatment, for example for hormone replacement aftermenopause. Tissues that can benefit from protection in view of thepresence of ERβ-receptors are prostate, testes (human), lung, colon andendometrium. In particular, endometrium proliferation can be reduced byERβ antagonists of the invention.

The compounds can be produced by various methods known in the art oforganic chemistry in general. More specifically the routes of synthesisas illustrated in the schemes and examples can be used.

-   -   R is protecting group

With reference to scheme 1, the benzofluorene (Z=CH₂) and thebenzanthracene (Z=CH₂CH₂) skeleton can be assembled in an identicalmanner. In step A adequately substituted indanones or tetralones aretreated with CS₂ under appropriate basic conditions to introduce adithioketene function (in fact serving as a carboxylate equivalent),after which procedure reaction with an organometallic derivative of asubstituted benzylhalide (preferably a Grignard derivative) in step B,followed by alcoholysis (step C) leads to an α,β-unsaturated ester. Atthis stage an acid catalyzed cyclization (step D) immediately leads tothe phenolic benzofluorene (or benzanthracene). Conversion of this intoa reactive intermediate (like triflate) in step E allows theintroduction of the desired functionalities (like aryl groups,carboxylates etc) by means of known organometallic techniques.

If the mentioned α,β-unsaturated ester is first hydrogenated in step Fprior to cyclization (step G), the indicated ketones become available.They may be easily converted into the aromatic iodide in step H. These,under circumstances may be more reactive than the afore-mentionedtriflates and provide valuable alternatives for functionalization (stepI in scheme 2).

Alkylation on the 11 [H] position can be performed in two separatesteps, using appropriate bases to generate anions at the methyleneposition of the 11[H]benzofluorene skeleton followed by treatment withalkylating agents. Deprotection of the silyl protecting group, followedby standard functional group transformations provides the desiredamines.

Ester prodrugs can be made by esterification of compounds with freehydroxyl groups by reaction with appropriate acyl chlorides in pyridine.Free dihydroxy compounds having formula 1 can be made by hydrolysis ofether precursors.

The present invention also relates to a pharmaceutical compositioncomprising the non-steroidal compound according to the invention mixedwith a pharmaceutically acceptable auxliary, such as described in thestandard reference Gennaro et al, Remmington: The Science and Practiceof Pharmacy, (20th ed., Lippincott Williams & Wilkins, 2000, seeespecially Part 5: Pharmaceutical Manufacturing). Suitable auxiliariesare made available in e.g. the Handbook of Pharmaceutical Excipients(2^(nd) Edition, Editors A. Wade and P. J. Weller; AmericanPharmaceutical Association; Washington; The Pharmaceutical Press;London, 1994). The mixture of the compounds according to the inventionand the pharmaceutically acceptable auxiliary may be compressed intosolid dosage units, such as pills, tablets, or be processed intocapsules or suppositories. By means of pharmaceutically suitable liquidsthe compounds can also be applied as an injection preparation in theform of a solution, suspension, emulsion, or as a spray, e.g. nasalspray. For making dosage units, e.g. tablets, the use of conventionaladditives such as fillers, colorants, polymeric binders and the like iscontemplated. In general any pharmaceutically acceptable additive whichdoes not interfere with the function of the active compounds can beused. The compounds of the invention may also be included in an implant,a vaginal ring, a patch, a gel, and any other preparation for sustainedrelease.

Suitable carriers with which the compositions can be administeredinclude lactose, starch, cellulose derivatives and the like, or mixturesthereof used in suitable amounts.

Furthermore, the invention relates to the use of the compounds for themanufacture of a medicine for an estrogen receptor related treatment,characterised in that the treatment is to antagonise the ERβ. In thecontext of the present use the antagonism does not have to be complete,but also includes partial antagonism.

Furthermore, the invention relates to the use of the non-steroidalcompound according to the invention for the manufacture of a medicamentfor estrogen-receptor related treatments and treatment ofestrogen-receptor related disorders such as peri- and/or post-menopausalcomplaints. Thus the invention also pertains to the medical indicationsof peri- and/or post-menopausal (climacteric) complaints andosteoporosis, i.e. a method of treatment in the field of hormonereplacement therapy (HRT), comprising the administration to a patient,being a woman, of a compound as described hereinbefore (in a suitablepharmaceutical dosage form).

Further, the invention relates to the use of the non-steroidal compoundaccording to the invention in the manufacture of a medicament havingcontraceptive activity. Thus the invention also pertains to the medicalindication of contraception, i.e. a method of contraception comprisingthe administration to a subject, being a woman or a female animal, of aprogestogen and an estrogen as is customary in the field, wherein theestrogen is a compound as described hereinbefore (in a suitablepharmaceutical dosage form).

Finally the invention relates to the use of the non-steroidal compoundfor the manufacture of a medicament having selective estrogenic and/oranti-estrogenic activity, such a medicament being generally suitable inthe area of HRT (hormone replacement therapy).

The dosage amounts of the present compounds will be of the normal orderfor estrogenic compounds, e.g. of the order of 0.01 to 100 mg peradministration.

The invention is further illustrated hereinafter with reference to someunlimitative examples and the corresponding formula schemes referred to.Compounds are identified by numbers (in bold letter type) with referenceto the corresponding numbers in the schemes. Abbreviations used in theschemes: Me is methyl, Bn is benzyl, ph is phenyl, aryl represents thesubstituted phenyl as in formula 1.

EXAMPLES Example 1

Preparation of Precursor 10-iodo-2,8-dihydroxy-11H-benzo[b]fluorene(4b).

59 ml 4-methoxybenzyl-magnesium chloride (0.2 M in diethyl ether) wasadded to 1 [J. V. Ram and M. Nath, Indian J. Chem. Sect. B; 34, 416-422(1995)] (11.6 mmol) in 70 ml THF at 0° C. and the reaction mixture wasstirred for 0.5 hour at 20° C. The mixture was poured into saturated aq.NH₄Cl, extracted with diethyl ether and dried over MgSO₄. Afterevaporation of the solvent the crude product was purified bychromatography on silica gel (heptane/ethyl acetate). The pure fractionswere concentrated and the material obtained was taken up in 95 mlmethanol and treated with BF₃.Et₂O (28 mmol). After 0.5 hour thetemperature was raised to 65° C. and after 0.5 hour the reaction mixturewas poured into water, extracted with CH₂Cl₂ and the organic layerwashed with NaHCO₃ (aq). The extract was dried over MgSO₄, concentratedand the residue was recrystallised from methanol to afford pure 2 in 45%yield (Rf=0.48 heptane/ethyl acetate (3:2)).

A mixture of 2 (5 mmol) and palladium on carbon (10% Pd (w/w), 300 mg)in 120 ml ethanol/acetic acid (5:1) was stirred under an atmosphere ofhydrogen for 1 hour. The catalyst was removed by filtration and thefiltrate was concentrated.

The residue was dissolved in methanesulfonic acid and stirred at 90° C.for 15 minutes after which the mixture was poured into ice water andextracted with ethyl acetate. The organic layer was washed withNaHCO₃(aq) and dried over MgSO₄. Chromatography on silica gel(heptane/ethyl acetate) gave pure 3 in 85% yield. (Rf=0.49 heptane/ethylacetate (2:1)); MP 96-98° C.

The compound 3 (0.34 mmol) was dissolved in ethanol and 1 ml hydrazinemonohydrate was added. After 4 hours refluxing, water was added and thehydrazone was extracted with CH₂Cl₂. The organic layer was washed withwater, dried and concentrated. The residue was taken up in 1.5 mltriethylamine and 0.2 g iodine in 0.7 ml THF was added at 0° C. After 1hour the reaction mixture was diluted with toluene, poured into icewater and extracted with toluene. The organic layer was washed with 1NHCl and saturated NaHCO₃(aq), dried over MgSO₄ and concentrated. Theresidue was dissolved in 8 ml m-xylene/toluene (2:1) palladium on carbon(10% w/w, 100 mg) was added and the mixture was heated at 125° C. for 2hours. After cooling the catalyst was filtered off, the filtrate wasconcentrated and the residue was purified on silica gel (heptane/ethylacetate). The appropriate fractions were collected and concentrated togive pure 4a. Compound 4a was dissolved in 30 mL CH₂Cl₂ and treated withBBr₃ (3.5 mmol). After 1 hour another 2.1 mmol of BBr₃ was added. After1.5 hours the mixture was carefully poured into sat. NaHCO₃ (aq) andextracted with ethyl acetate. The organic layer was dried over MgSO₄ andconcentrated. Chromatography on silica gel (toluene/ethyl acetate)afforded pure 4b in 62% yield. (Rf=0.50 toluene/ethyl acetate (4:1));ESI-MS: M+H=375.2, M−H=373.0.

General Procedure to Prepare Compounds 5a-v(10-aryl-2,8-dihydroxy-11H-benzo[b]fluorenes)

(Reference to Scheme 3)

A mixture of 10-iodo-benzofluorene derivative 4 (27 μmol), 3 mgPd₂(dba)₃, 0.2 M Na₂CO₃(aq), 30 μmol arylboronic acid and 1 ml2-methoxy-ethanol was heated for 5 hours at 55° C. Ethyl acetate andwater were added to the reaction mixture and the organic layer wasseparated, dried over MgSO₄ and concentrated. The residue was purifiedon silica gel (toluene/ethylacetate) to give pure 5a-v (yields 14-52%).

Compound ARYL Yield (%) [M − H] 5a 4-chlorophenyl 37 [M − H] = 357.2 5b2-naphthyl 44 [M − H] = 373.2 5c 3-methoxyphenyl 32 [M − H] = 353.4 5d3-trifluoromethylphenyl 54 [M − H] = 391.3 5e 4-methylphenyl 42 [M − H]= 337.4 5f 3-chloro-4-fluorophenyl 40 [M − H] = 375.2 5g3,4-methylenedioxophenyl 49 [M − H] = 367.4 5h 4-phenylphenyl 55 [M − H]= 399.4 5i 2-benzothiazole 30 [M − H] = 379.4 5j 3-fluorophenyl 27 [M −H] = 341.4 5k 4-methoxyphenyl 27 [M − H] = 353.4 5l 4-fluorophenyl 52 [M− H] = 341.4 5m 3,4-dichlorophenyl 14 [M − H] = 390.8 5n 3-chlorophenyl37 [M − H] = 357.0 5o 4-trifluoromethylphenyl 22 [M − H] = 391.4 5p3-methylphenyl 21 [M − H] = 337.2 5q 3-isopropylphenyl 40 [M − H] =365.0 5r 4-trifluoromethyloxyphenyl 41 [M − H] = 407.2 5s3-fluoro-4-phenylphenyl 22 [M − H] = 417.0 5t 4-methylthiophenyl 32 [M −H] = 371.2 5u 2-trifluoromethylphenyl 20 [M − H] = 391.0 5v Phenyl 25 [M− H] = 323.2

Example 2

Compound 7a-d

A mixture of 4b (0.94 mmol), potassium carbonate (3.0 mmol) and benzylbromide (2.1 mmol) in acetone (10 ml) was refluxed overnight after whichthe mixture was poured into water and extracted with ethyl acetate. Theorganic layer was dried over MgSO₄, concentrated and purified bychromatography on silica gel (heptane/ethyl acetate).

The purified product (0.43 mmol) was taken up in 2-methoxyethanol (16ml) and Pd₂(dba)₃ (36 μmol), 3-hydroxyphenylboronic acid pinacolester(0.45 mn and Na₂CO₃ (2M in water, 2 ml) were added. The mixture wasstirred for 30 minutes at 60° C., poured into water and extracted withethyl acetate. The organic layer was dried over MgSO₄, concentrated andpurified by chromatography on silica gel (CH₂Cl₂/methanol) to give pure6 in 56% yield=0.34 (heptane/ethyl acetate (7:3)).

A mixture of 6 (48 μmol), 1-(2-chloroethyl)pyrrolidine hydrochloride (76μm and cesium carbonate (0.15 mmol) in acetonitrile (2 ml) was stirredfor 3 hr at 50° C. The mixture was poured into water and extracted withethyl acetal the organic extract was dried over MgSO₄, the solventevaporated and the residue was purified by chromatography on silica gel(CH₂Cl₂/methanol). The pure fractions were concentrated and the materialobtained was dissolved in ethyl acetate (3 ml). Pd/C (10% w/w, 25 mg)and 3 drops of acetic acid were added and the mixture was stirred underan atmosphere of hydrogen for 5 hours. The catalyst was removed byfiltration and the filtrate was concentrated. The residue was purifiedby chromatography on silica gel (CH₂Cl₂/methanol yield pure 7a in 22%yield. Rf=0.14 (CH₂Cl₂/methanol (9:1)), ESI-MS: M+H 438.4, M−H=436.2.

Compound 7b

Compound 7b was prepared from 6 in 5% yield, in the same fashion asdescribed for the preparation of 7a, using 2-dimethylaminoethyl chloridehydrochloride (Rf=0.18 CH₂Cl₂/methanol (9:1)); ESI-MS: M+H=412.4,M−H=410.4.

Compound 7c

Compound 7c was prepared from 6 in 32% yield, in the same fashion asdescribed for the preparation of 7a, using 1-(2-chloroethyl)morpholinehydrochloride instead of 1-(2-chloroethyl)pyrrolidine hydrochloride(Rf=0.21 CH₂Cl₂/methanol (9:1)); ESI-MS: M+H=454.4, M−H=452.2.

Compound 7d

Compound 7d was prepared from 6 in 65% yield, in the same fashion asdescribed for the preparation of 7a, using 2-diethylaminoethyl chloridehydrochloride instead of 1-(2-chloroethyl)pyrrolidine hydrochloride(Rf=0.17 CH₂Cl₂/methanol (9:1)); ESI-MS: M+H=440.4, M−H=438.2.

Compound 7e

Compound 7e was prepared from 6 in 18% yield, as described for thepreparation of 7a, using 1-(2-chloroethyl)piperidine hydrochlorideinstead of 1-(2-chloroethyl)pyrrolidine hydrochloride (Rf=0.15CH₂Cl₂/methanol (9:1)); ESI-MS: M+H=452.4, M−H=450.2.

Compound 9

A mixture of 4a (0.30 mmol), Pd₂(dba)₃ (0.40 μmol),4-hydroxyphenylboronic acid (0.30 mmol) and sodium carbonate (2 M inwater, 4 ml) in 12 ml 2-methoxyethanol was stirred at 60° C. After 30minutes the mixture was poured into water and extracted with ethylacetate. The organic extract was dried over MgSO₄, concentrated andpurified by chromatography on silica gel (toluene/ethyl acetate) to give8 in 65% yield. Rf=0.24 (toluene/ethyl acetate (8:2)).

Compound 8 (0.16 mmol) was dissolved in toluene (3 ml). Sodium hydride(0.4 mmol) and 1-(2-chloroethyl)piperidine hydrochloride (0.2 mmol) wereadded and the mixture was refluxed for 3.5 hours. The reaction mixturewas poured into water and extracted with ethyl acetate. The organicextract was dried over MgSO₄, concentrated and purified bychromatography on silica gel (toluene/methanol).

The pure fractions were collected and concentrated, the materialobtained (46 μmol) was dissolved in CH₂Cl₂ and treated with ethanethiol(0.62 mmol) and aluminum chloride (95 μmol) at RT. After 16 hours thedark red mixture was poured into water and extracted with ethyl acetate.The organic extract was dried over MgSO₄, concentrated and purified bychromatography on silica gel (CH₂Cl₂/methanol) to give 9 in 22% yield.Rf=0.23 (toluene/methanol (85:15)), ESI-MS: M+H=452.4, M−H=450.2.

Example 3

Compound 12a

A mixture of 3-hydroxyphenylboronic acid pinacolester 10a (0.68 mmol),cesium carbonate (0.68 mmol) and 1-bromo-3-chloropropane (0.80 mmol) inacetonitrile (3 ml) was stirred overnight at RT. Additional cesiumcarbonate (0.31 mmol) and 1-bromo-3-chloropropane (0.4 mmol) were addedand the mixture was stirred overnight at 60° C. The mixture was pouredinto water and extracted with CH₂Cl₂. The CH₂Cl₂-layer was dried overMgSO₄, concentrated and purified by chromatography on silica gel(toluene/ethyl acetate). The purified product was dissolved inpiperidine and stirred for 48 hours at 45° C. The solid material(piperidine.HCl) was filtered off and the filtrate was concentrated togive 11a in 88% yield. Rf=0.05 (toluene/ethyl acetate (4:1)).

A mixture of 4b (67 μmol), 11a (86 μmol), PdCl₂(dppf₂ (5 μmol) andsodium carbonate (2 M in water, 0.25 ml) in 2.5 ml 2-methoxyethanol wasstirred at 90° C. After 2 hours the mixture was poured into water andextracted with ethyl acetate. The organic extract was dried over MgSO₄,concentrated and purified by chromatography on silica gel(CH₂Cl₂/methanol). The appropriate fractions were collected andconcentrated, the material obtained was recrystallised from CHCl₃ togive 12a in 38% yield. Rf=0.42 (CH₂Cl₂/methanol (85:15)).

Compound 12b

A mixture of 4-hydroxyphenylboronic acid pinacolester 10b (0.68 mmol),potassium hydroxide (2.1 mmol) and 1-bromo-3-chloropropane (2.8 mmol) inmethanol (2 ml) was refluxed for 24 hours. The mixture was poured intowater and extracted with ethyl acetate. The organic extract was driedover MgSO₄, concentrated and purified by chromatography on silica gel(toluene/ethyl acetate). The purified product was dissolved inpiperidine and stirred overnight at 50° C. The solid material(piperidine.HCl) was filtered off and the filtrate was concentrated togive 11b in 80% yield. Rf=0.10 (toluene/methanol (9:1)).

Compound 12b was prepared from 4a and 11b in 20% yield, in a similarfashion as described for the preparation of 12a. Rf=0.42(CH₂Cl₂/methanol (85:15)), ESI-MS: M+H=466.4, M−H=464.6

Compound 12c

Compound 12c was prepared from 10a in 25% yield, as described for thepreparation of 12a, using 1-bromo-4-chloro-butane instead of1-bromo-3-chloro-propane. Rf=0.21 (CH₂Cl₂/methanol (8:2)), ESI-MS:M+H=480.6, M−H=478.2

Compound 12d

To mixture of 1,4-diiodobutane (5 mmol) and cesium carbonate (0.68 mmol)in acetonitrile (2 ml) was portionwise added 4-hydroxyphenylboronic acidpinacolester 10b (0.68 mmol) at 40° C. After 2.5 hours water was addedand the mixture was extracted with ethyl acetate. The organic layer wasdried over MgSO₄, concentrated and purified by chromatography on silicagel (heptane/toluene). The purified product was dissolved in piperidineand stirred at RT for 2 hours. The solid material (piperidine.HI) wasfiltered off and the filtrate was concentrated to give 11d in 32% yield.Rf=0.55 (toluene/methanol (8:2)).

Compound 12d was prepared from 4b and 11d in 13% yield, in a similarfashion as described for the preparation of 12a. Rf=0.22(CH₂Cl₂/methanol (8:2)), ESI-MS : M+H 480.4, M−H=478.2

Example 4

Compound 14

A mixture of 2.03 mmole of 1,3-dibromopropane and 1.02 mmole ofpotassium carbonate in 10 ml of acetone was warned to 40° C. To thissolution 0.51 mmole of 13 in 10 ml of acetone was added dropwise and thereaction mixture was stirred at 40° C. for 23 hours. An additional ismixture of 2.03 mmole of 1,3-dibromopropane and 1.02 mmole ofpotassiumcarbonate in 5 ml of acetone was added and the reaction mixturestirred for 4 hours at reflux temperature. The reaction mixture wastaken up in ethyl acetate and water, washed with water and saturatedNaCl solution, dried over MgSO₄ and concentrated. The crude product waspurified by chromatography on silica gel (heptane/ethyl acetate) to givepure 14 in 65% yield. Rf=0.64 (heptane/diethylether (7:3))

Compound 15a

82 μmole of 14 was dissolved in 6 ml of dry CH₂Cl₂. 327 μmole ofBF₃.S(CH₃)₂ was added and the solution was stirred at room temperaturefor 16 hours. The reaction mixture was taken up in ethyl acetate, washedwith water and saturated NaHCO₃ solution, dried over MgSO₄ andconcentrated. The crude product was purified by chromatography on silicagel (CH₂Cl₂/methanol) to give pure 15a in 93% yield. Rf=0.47(CH₂Cl₂/methanol (9:1))

Compound 15b

22 μM of bromide 14 was refluxed for 1.5 hours with 100 μM LiAlH₄ inTHF. Water and ethyl acetate were added to the reaction mixture and theorganic layer was separated, dried over MgSO₄ and concentrated. Theresidue was purified on silicagel (methylene/methanol) to give pure3′-O-propyl compound 15b in yield 37%. Rf=0.40 (heptane-ethyl acetate7:3).

Compound 15c

54 μM of compound 13 was reacted with 1.7 mM 1-bromo-3-phenylpropane inthe presence of 1.7 mM K₂CO₃ in 3 ml acetone at room temperature. After24 hours the salts were removed by filtration. The filtrate wasconcentrated and redissolved in methylene chloride. The mixture wasextracted with water, dried over MgSO₄ and concentrated. The residue waspurified by chromatography on silica gel (heptane/ethylacetate).(yield=88%).

47 μM of the resulting product was demethylated with 1.9 mM (CH₃)₃S.BF₃in CH₂Cl₂ for one night. Ethyl acetate and water were added to thereaction mixture and the organic layer was separated, dried over MgSO₄and concentrated. The residue was purified on silica gel(heptane/ethylacetate) to give pure compound 15c in yield=57%. Rf 0.7(heptane ethyl acetate 8:2)

Example 5

2,8-dimethoxy-10-hydroxy-11H-benzo[b]fluorene (Compound 16)

The compound 2,8-dimethoxy-10-hydroxy-11H-benzo[b]fluorene (Compound 16)1 was prepared from its corresponding ester as explained above for step4 in scheme 1. An amount of 3 g of the corresponding unsaturated esterwas added in small portions over a few minutes to 30 ml ofmethanesulphonic acid at 60° C. After stirring for ½ hr the cyclizationwas complete. The mixture was then poured onto ice water and stirred foran additional ½ hr. The product was filtered, washed with water andthoroughly dried over P₂O₅, to give 2.2 gr of compound 16.

R_(f)0.38 (heptane/eth. ac. 7/3). NMR (DMSO) 3.82, 3.88 (2×3H, s, OCH₃),3.95 (s, 2H, CH₂), 9.57 (s, 1H, OH), 6.97, 7.11, 7.20, 7.55, 7.51, 7.75,7.80 (7H's, aromatic protons)

5-chloro-2,8-dimethoxy-10-hydroxy-11H-benzo[b]fluorene (Compound 17)

To a solution of 800 mg of compound 16 in 10 ml of DMF was added 850 mgof 2,2,3,4,5,6-hexachlorocyclohexa3,5-diene in small portions over 5minutes. The mixture was stirred for 1 hr and then poured into 50 ml ofwater. The dark reaction product was extracted with ethyl acetate andpurified by chromatography over silica gel (heptane/ethyl acetate aseluent), to provide 380 mg of 17 as a brown solid; R_(f) 0.38(hept./ethyl ac. 6/4), R_(f) (starting material) 0.44. NMR (DMSO) 3.85,3.92 (2×s, 6H, OCH₃) 4.03 (s, 2H, CH₂), 7.03, 7.30, 8.13, 8.38 (2×AB,4H, Ar—H), 7.25, 7.61 (2×br.s, 2H, Ar—H).

Compound 18

To a solution of 900 mg of 17 in 8 ml of pyridine was added at 0° C. 700μl of trifluoromethanesulphonic anhydride. Stirring was performed for 1hr at RT followed by pouring into water and additional stirring for 15min. followed by filtration of the crude product. Purification wasachieved by chromatography over silicagel, to provide 800 mg of triflate18; Mp 165-168° C. NMR (CDCl₃) 3.90, 3.96 (2×s, 6H, OCH₃), 4.18 (s, 2H,CH₂), 7.0, 7.09, 7.29, 7.35, 8.11, 8.47 (6H, Ar—H).

Compound 19

A mixture of 210 mg of triflate 18, 220 mg of3-hydroxyphenyl-pinacolborane, 200 mg of K₃PO₄, 15 mg of As(PPh)₃, 15 mgof PdCl₂.PPh₃, 0.5 ml of water and 5 ml of dioxane was heated at 100° C.for 1.5 hr under a nitrogen atmosphere. The reaction was poured intowater and extracted with ethyl acetate. Chromatography of the resultingmaterial provided 215 mg of 19 as an amorphous product; R_(f) 0.35(hept./ethyl ac. 7/3), Mp 184-185° C. NMR (CDCl₃) 3.74, 3.87 (s, 6H,OCH₃), 3.80 (s, 2H, CH₂), 6.82-7.0 (m, 6H, Ar—H), 7.25, 7.40, 8.38, 8.53(4H, Ar—H).

Compound 20

A mixture of 200 mg of 19, 500 mg of powdered K2CO3, 1.25 ml of1,3-dibromopropane and 10 ml of acetonitrile was heated at 55° C. for 3hr. The reaction was diluted with water and extracted with ethylacetate. The crude product was purified by chromatography on silica gel(hept./ethyl acetate), to provide 220 mg of 20; R_(f) 0.63(hept./eth.ac. 7/3); NMR (CDCl₃) 3.65 (t, 2H, CH₂Br), 2.33 (m, 2H, CH₂),4.13 (t, 2H, CH₂₀), 3.78 (s, 2H, CH₂).

Compound 21

To a solution of 220 mg of 20 in 7 ml of methylenechloride was added 1.5ml of BF3.dimethylsulfde complex. The mixture was stirred untilcompletion of the reaction (5 hr). The reaction was poured into waterand the product extracted with ethyl acetate. Chromatography provided210 mg of 21 as a colorless amorphous material; R_(f) 0.25(hept./eth.ac. 7/3). NMR (CDCl₃) 3.67 (t, 2H, CH₂Br), 2.33 (m, 2H, CH₂),4.15 (t, 2H, CH₂O), 3.77 (s, 2H, CH₂).

Compound 22

A mixture of 70 mg of 21, 0.3 ml of 1,2,5,6-tetrahydropyridine and 3 mlof acetonitrile was heated at 55° C. for ½ hr. The mixture was thenpoured onto 5% NaHCO₃ and extracted with ethyl acetate. The product waspurified by passing through a short silica column (CH₂Cl₂/CH₃OH). Theproduct thus obtained was converted into a HCl salt by treatment of asolution the free base in methanol/ether with 1M HCl/ether. Thehydrochloride salt thus obtained was freeze-dried from water to obtain45 mg of amorphous 22. NMR (DMSO) 9.77, 9.82 (2×s, 2H, OH's), 5.70 and5.88 (2×m, 2H, tetrahydropyridine), 8.32, 8.20, 7.52, 7.21, 7.08, 6.98,6.87 (10, aromatic H's).

Example 6

3,9dimethoxy-7-hydroxy-5,6-dihydro-benz[a]anthracene (compounds 23) and12chloro-3,9-dimethoxy-7-hydroxy-5,6-dihydro-benz[a]anthracene (compound24)

The compound 3,9-dimethoxy-7-hydroxy-5,6-dihydro-benz[a]anthracene(compounds 23) was prepared analogously to compound 16 in example 5. Toa solution of 600 mg of 23 in 10 ml of DMF was added in portions 600 mgof 2,3,4,4,5,6-hexachlorocyclohexa-2,5-dien-1-one. The mixture was thenstirred at 40° C. for 4 hr. Then the reaction was poured into water andthe product extracted with ethyl acetate. The crude material was passedthrough a silicagel column (hept./eth.ac.) and finally triturated withheptane-diisopropyl ether to provide 280 mg of 24 as orange crystals; Mp140-141° C., R_(f) 0.28 (hept./eth.ac. 7/3) starting material R_(f)0.30.

Compound 25

To a solution of 300 mg of 24 in 3 ml of pyridine was added 200 μl oftriflic anhydride. The mixture was stirred for 1 hr at rt, and thenpoured into water and extracted with ethyl acetate. The product waspurified over silica gel and afforded 220 mg of 25 as a white solid; Mp122-124; R_(f) 0.70 (hept./ethyl ac. 7/3).

Compound 26

A mixture of 210 mg of 25, 220 mg of 3-hydroxyphenylpinacolborane, 200mg of K₃PO₄ 15 mg of (PPh₃)As, 15 mg of PdCl₂(PPh₃)₂, 0.5 ml of waterand 5 ml of dioxane was heated at 100° C. for 1.5 hr. The mixture wasthen poured into water and extracted with ethyl acetate. Chromatographyover silica gel provided 215 mg of 26 as an oil; R_(f) 0.28 (hept./ethylacetate 7/3). NMR (DMSO) 2.56 (4H, CH₂CH₂), 3.67, 3.80 (2×s, 6H, OCH₃),8.32, 8.18, 7.33, 6.93, 6.70 (10H, Ar—H's), 9.64 (s, 1H, OH).

Compound 27

A mixture of 215 mg of 26, 500 mg of K₂CO₃, 1.2 ml of 1,3-dibromopropaneand 10 ml of acetonitrile was heated at 55° C. for 2.5 hr. The reactionwas then poured in water and extracted with ethyl acetate.Chromatography provided 220 mg of 27 as a colorless oil; R_(f) 0.60(hept./ethyl acetate 7/3). NMR (CDCl₃) 2.60 (m, 4H, CH₂CH₂), 2.30 (m,2H, CH₂), 3.60 (t, 2, CH₂Br), 4.13 (t, 2H, CH₂O), 3.72, 3.87 (2×s, 6H,OCH3).

Compound 28

To a solution of 190 mg of 27 in 7 ml of methylenechloride was added 1.5ml of BF₃.dimethylsulfide complex. After stirring at rt for 4 hr themixture was poured onto water and extracted with ethyl acetate.Chromatography of the crude product gave 150 mg of essentially pure 28;R_(f) 0.20 (hept./ethyl ac. 7/3); NMR (DMSO) 2.27 (m, 2H, CH₂), 2.50 (m,4H, CH₂CH₂), 3.68 (t, 2H, CH₂Br), 4.12 (t, 2H, CH₂O), 9.68, 9.82 (2×s,2, OH).

Compound 29

A mixture of 60 mg of 28, 0.4 ml of pyrrolidine and 3 ml of acetonitrilewas stirred at 50° C. for ½ hr. The mixture was then poured into 5%NaHCO3 and extracted with ethyl acetate. The product was purified bypassing through a short silica column (CH₂Cl₂/CH₃OH as eluent) and thenconverted into a HCl salt by treatment with 1M HCl/ether. The resultinghydrochloride was freeze dried from water to give 35 mg of 29; R_(f)0.20 (CH₂Cl₂/CH₃OH/HOAc 90/10/1); NMR (DMSO) 9.70 and 9.82 (2×s, 2H,OH's), 8.22, 8.05, 7.48 7.17, 7.06, 6.88, 6.84, 6.76, 6.70, 6.62 (m,10H, Ar—H's), 4.10 (t, 2H, CH₂O).

Example 7

Compound 30

A mixture of 300 mg of compound 13, 900 mg of powdered K₂CO₃, 2 ml of1,2-dibromopropane and 8 ml of acetonitrile was heated at 55° C. for 16hr. The reaction was diluted with water and extracted with ethylacetate. S The crude product was purified by chromatography on silicagel (hept./ethyl acetate), to provide 310 mg of 30; R_(f) 0.50(hept./eth.ac. 7/3); NMR (CDCl₃) 3.67 (t, 2H, CH₂Br), 4.35 (t, 2H,CH₂O), 3.79 (s, 2H, CH₂), 3.75, 3.87 (2×s, 6H, OCH₃).

Compound 31

To a solution of 310 mg of 30 in 6 ml of methylenechloride was added 2ml of BF₃.dimethylsulfide complex. The mixture was stirred untilcompletion of the reaction (5 hr). The reaction was poured into waterand the product extracted with ethyl acetate. Chromatography provided290 mg of 31 as a colorless amorphous material; R_(f) 0.19(hept./eth.ac. 7/3). NMR (CDCl₃) 3.67 (t, 2H, CH₂Br),), 4.35 (t, 2H,CH₂O), 3.76 (s, 2H, CH₂).

Compound 32

A mixture of 60 mg of 31 0.3 g of 2-pyrimidinylpiperazine and 2 ml ofacetontrile was heated at 50° C. for 16 hr. The mixture was then dilutedwith water and the product extracted with ethyl acetate. The organicmaterial was passed through a short silica column (a gradient ofCH₂Cl₂/CH₃OH as eluent), to provide essentially pure 32 as the freebase. This was dissolved in a small amount of ethyl acetate and treated25 with 1M HCl in ether to give the HCl salt. This was freeze dried fromwater to provide 48 mg of amorphous HCl salt of 32. R_(f) 0.82(CH₂Cl₂—CH₃OH— acetic acid 9/1/0,1); NMR (DMSO) 4.50 (m, 2H, CH₂O),6.76, 6.84, 6.88, 6.96, 7.05, 7.09, 7.16, 7.21, 7.55, 8.21, 8.32, 8.44(resp. 1H, 1H, 1H, 1H, 1H, 1H, 1H, 1H, 1H, 1H, 1H, 2H's; Ar—H's).

Example 8

Compound 33

To 610 mg of NaBH4 in 20 ml of THF was added dropwise 2.7 ml of aceticacid. The mixture was allowed to stirr for 1 hr. A mixture of 1 ml ofp-methoxybenzaldehyde and 3-amino-propanol in 2 ml of THF was stirredfor ½ hr and then added to the aforementioned hydride solution. This wasstirred overnight. The reaction was poured into water and acidified with2N HCl. The acidic phase was washed with ethyl acetate and then treatedwith 2N NaOH to make the mixture slightly basic. Extraction with ethylacetate, followed by washing, drying and concentration provided 880 mgof 33; NMR (CDCl₃) δ 6.88 and 7.23 (AB, 4H, Ar—H's); 1.72 (m, 2H), 2.90(m, 2H), 3.74 (s, 2, ArCH₂N—), 3.82 (s and d, 5H, OCH₃ and CH₂OH)

Compound 34

A mixture of 120 mg of 15a and 250 mg of 33 in 2 ml of acetonitrile washeated at 60° C. for 16 hr. The reaction was poured into 5% NaHCO₃ andextracted with ethyl acetate. The product was purified by passingthrough a short silica column (dichloromethane-methanol). The productthus obtained was taken up in 2 ml of ether and treated with 1 eq of 1MHCl in ether. The precipitate was isolated by centrifugation of thesupernatant and dissolved in water and freeze dried, to give 110 mg of34.

NMR δ (DMSO) 9.50 (2×s, phenolic OH's), 1.87 (m, 2H), 2.20 (m, 2H), 3.08(m, 2H), 3.18 (m, 2H), 3,46 (m, 2H) 4.21 (m, 2H), 3.68 (m, 2H), 3.70 (s,3H, OCH₃) 4.10 (m, 2H); R_(f) 0.43 (dichloromethane/methanol 9/1).

Example 9

Compound 37

To a solution of 3 gr of silylether protected phenol 36 in 25 ml of dryTHF (under N₂) was added at −20° C., 12.5 ml of 1M of LiHMDS solution inTHF. The blue reaction mixture was stirred for an additional 45 min,after which period 1 ml of methyliodide was added The mixture wasstirred for an additional ½ hr and then poured onto 100 ml of water andextracted with ethyl acetate. After washing, drying and concentration3.2 gr of 37 were isolated as amorphous solid in essentially pure form;NMR (CDCl₃): δ 0.22 (d, 6H, (CH₃)₂—Si), 1.01(s, 9H, tBuSi), 1.03(d, 3H,CHCH₃), 3.71 and 3.73 (3H, 2×OCH3 rotamers) 3.87 and 3.88 (3H, 2×OCH3rotamers), 4.15 (m, 1H, CH(CH₃)) 6.92-8.0 (11 Ar—H's); TLC: R_(f) 0.69(hept/ethyl acetate 7/3, the same value as the starting material).

Compound 38

A solution of 600 mg of 37 in 5 ml of dry THF was treated with 1 ml of1.6M BuLi in hexane at −60° C. The reaction mixture was stirred for 1 hrat −30° C. and then treated with 1 ml of methyliodide. The mixture wasstirred for ½ hr at −30° C. and then warmed to room temperature andquenched by addition of water. The product was extracted with ethylacetate and the organic phase was washed, dried and concentrated, toprovide 580 mg of 38 in essentially pure form; R_(f.)0,70 (hept/ethylacetate 7/3). (NMR (CDCl₃): δ 0.20 (2s, 6H, (CH₃)₂—Si); 0.98(s, 9H,(CH₃)₃Si), 1.34 (2×s, 6H, C(CH₃)₂), 3.63 (s, 3H, OCH₃), 3.87(s, 3H,OCH₃)

Compound 39

To a solution of 600 mg of 38 in 2 ml of THF was added 1.6 ml of 1M+TBAF in THF. The mixture was stirred for 15 minutes and then treatedwith 25 ml of sat. NH₄Cl solution and extracted with ethyl acetate. Theproduct thus obtained was purified by passing through a short silicacolumn, to provide 400 mg of 39 as a white solid; Mp 198-200° C.; NMR(CDCl₃) δ 1.35 and 1.30 (2s, 6H, C(CH₃)₂), 3.66, 3.87(6H, 2×OCH3), 4.85(broad s, OH); R_(f) 0.37 (hept/ethyl acetate 7/3).

Compound 40

A mixture of 350 mg of 39, 750 mg of dry K₂CO₃, 2.3 ml of1,3-dibromopropane and 10 ml of acetonitrile was stirred at 60° C. for 3hr. Then the reaction was poured into water and extracted with ethylacetate. The material thus obtained was purified by chromatography, togive 440 mg of colorless oil; R_(f)0.62 (hept/ethyl acetate 7/3). NMR(CDCl₃) δ 1.35 and 1.31 (2×s, 6H, C(CH₃)₂), 2.34 (m, 2H, —CH₂—), 3.62(t, 2H, CH₂Br), 4.14 (t, 2H, CH₂O), 3.66, 3.88 (2×s, 6H, OCH₃).

Compound 41

A solution of 440 mg of 40 in 5 ml of dichloromethane was treated with3.6 ml of BF₃.dimethylsulfide complex. The mixture was stirred overnightand then ice-water was added and the product extracted intomethylenechloride. Purification was achieved by passing through a shortsilica column (heptane/ethyl acetate as eluent), to provide 280 mg of41, Rf 0.19 (hept/ethyl acetate 7/3). NMR (CDCl₃)δ 1.33 and 1.30 (2×s,6H, CH₃), 2.33 (m, 2H, —CH₂—), 3.62 (t, 3H, CH₂Br), 4.14 (m, 2H, CH₂O),4.85 (broad s, 2H, OH's).

Compound 42

A solution of 65 mg of 41 in 1 ml of acetonitrile and 0.2 ml of1,2,3,6-tetrahydropyridine was heated at 60° C. for 3 hr. The mixturewas then poured into 10 ml of 5% aqueous NaHCO3 and extracted with ethylacetate. The product was passed through a short silica column (usingmethylenedichloride/methanol 95/5 as eluent). The purified material thusobtained was treated with one equivalent of 1M HCl in ether and thesolid thus obtained was freeze dried from water to give 46 mg of 42 asHCl salt. R_(f) 0.45 (CH₂Cl₂-methanol 9/1); NMR (DMSO) δ 9.43 and 9.53(2s, 2H, OH's), 8.05 (s, 1, ArH), 7.78 (1H, ArH), 7.70 (1H, ArH)7.49 (t,1H, ArH), 7.10 (dd, 1H, ArH) 7.02(dd, 1H, ArH), 6.88 (br s, 1H, ArH),6.91 (d, 1H, ArH)6.78(2H, m, ArH's), 6.42(d, 1H, ArH), 5.71 and5.90(2×br d, 2H, —CH═CH—), 1.23 (2s, 6H, CH₃).

Example 10

Biological Activity

Determination of competitive binding to cytoplasmic human estrogenreceptor α or β from recombinant CHO cells is used to estimate therelative affinity (potency ratio) of a test compound for estrogenreceptors present in the cytosol of recombinant Chinese hamster ovary(CHO) cells, stably transfected with the human estrogen receptor α (hERα) or β receptor (hERβ), as compared with 17β-estradiol (E₂).

The estrogenic and antiestrogenic activity of compounds is determined inan in vitro bioassay with recombinant Chinese hamster ovary (CHO) cellsstably co-transfected with the human estrogen receptor α (hERα) or βreceptor (hERβ), the rat oxytocin promoter (RO) and the luciferasereporter gene (LUC). The estrogenic activity (potency ratio) of a testcompound to stimulate the transactivation of the enzyme luciferasemediated via the estrogen receptors hERα or hERβ is compared with thestandard estrogen estradiol. The antiestrogenic activity (potency ratio)of a test compound to inhibit the transactivation of the enzymeluciferase mediated via the estrogen receptors hERα or hERβ by theestrogen estradiol is compared with the standard ICI 164.384 (=(7α,17β)-N-butyl-3,17-dihydroxy-N-methylestra-1,3,5(10)-triene-7-undecanamide).

Results (Table on Next Page)

ERβ Compound antagonism  5a +  5b +  5c +  5d +  5e ++  5f +  5g +  5h + 5I +  5j +  5k +  5l ++  5m ++  5n +  5o +  5p +  5q +  5r ++  5s + 5t +  5u ++  5v +  7a +++  7b +++  7c +++  7d +++  7e +++  9 ++ 12a +++12b ++ 12c +++ 12d + 14 ++ 17a +++ 17b ++ 17c ++ >5% (relative toICI): + >40%: ++ >100%: +++

1. A compound, or a pharmaceutically acceptable salt thereof, having theformula 1,

wherein: R^(e) and ′R^(e) are OH, optionally independently etherified oresterified; Z is —C(R⁴,R⁵)—, wherein R⁴ and R⁵ are independently(1C-2C)alkyl or form together a spiro(3C-5C)cycloalkyl; R¹ is H,halogen, CF₃, or (1C-4C)alkyl; R² and R³ are independently H, halogen,—CF₃, —OCF₃, (1C-8C)alkyloxy, aryloxy, aryl(1C-8C)alkyl,halo(1C-8C)alkyl, —O(CH₂)_(m),X, wherein X is halogen or phenyl andm=2-4; —O(CH₂)_(m)NR_(a)R_(b), —S(CH₂)_(m)NR_(a)R_(b) or—(CH₂)_(m)NR_(a)R_(b), wherein m=2-4 and wherein R_(a), R_(b) areindependently (1C-8C)alkyl, (2C-8C)alkenyl, (2C-8C)alkynyl, or aryl,which alkyl, alkenyl and aryl can optionally be substituted withhalogen, —CF₃, —OCF₃, —CN, —NO₂, —OH, (1C-8C)alkoxy, aryloxy, carboxyl,(1C-8C)alkylthio, carboxylate, (1 C-8C)alkyl, aryl, aryl(1C-8C)alkyl orhalo(1C-8C)alkyl; or R_(a) and R_(b) form a 3-8 membered ring structure,optionally substituted with halogen, —CF₃, —OCF₃, —CN, —NO₂, hydroxy,hydroxy(1C-4C)alkyl, (1C-8C)alkoxy, aryloxy, (1C-8C)alkylthio, carboxyl,carboxylate, (1C-8C)alkyl, aryl, aryl(1C-8C)alkyl, halo(1 C-8C)alkyl. 2.The compound, or a pharmaceutically acceptable salt thereof, accordingto claim 1, wherein R³ is H and R² is —O(CH₂)_(m)NR_(a)R_(b),—S(CH₂)_(m)NR_(a)R_(b) or —(CH₂)_(m)NR_(a)R_(b), wherein m=2-4 andwherein R_(a), R_(b) are independently (1C-8C)alkyl, (2C-8C)alkenyl,(2C-8C)alkynyl, or aryl, which alkyl, alkenyl and aryl can be optionallysubstituted with halogen, —CF₃, —OCF₃, —CN, —NO₂, —OH, (1C-8C)alkoxy,aryloxy, carboxyl, (1C-8C)alkylthio, carboxylate, (1C-8C)alkyl, aryl,aryl(1C-8C)alkyl or halo(1C-8C)alkyl; or R_(a) and R_(b) form a 3-8membered ring structure, optionally substituted with halogen, —CF₃,—OCF₃, —CN, —NO₂, hydroxy, hydroxy(1C-4C)alkyl, (1C-8C)alkoxy, aryloxy,(1C-8C)alkylthio, carboxyl, carboxylate, (1C-8C)alkyl, aryl,aryl(1C-8C)alkyl, halo(1C-8C)alkyl.
 3. The compound according to claim1, wherein R¹ is selected from H, halogen and CF₃.
 4. The compoundaccording to claim 2, wherein, R² is —O(CH₂)_(m)NR_(a)R_(b), whereinm=2-3 and R_(a), R_(b) are independently (1 C-5C)alkyl, (3C-5C)alkenyl,or aryl, which alkyl, alkenyl and aryl can be optionally substitutedwith OH or methoxy, or R_(a) and R_(b) form a 4-7 membered ringstructure selected from the list: azetidine, pyrrolidine, 3-pyrroline,piperidine, piperazine, tetrahydropyridine, morpholine, thiomorpholine,thiazolidine, homopiperidine, tetrahydroquinoline and6-azabicyclor[3.2.1]octane, which 4-7 membered ring structure canoptionally be substituted with OH, methoxy, acetyl, carboxylate,(1C-3C)alkyl, phenyl, benzyl, and phenylethyl.
 5. A method of treatingan estrogen receptor related disorder selected from the group consistingof contraception, benign prostate hypertrophy, osteoporosis, Alzheimer'sdisease and depression, comprising administering an effective amount ofthe compound according to claim 1 to a patient in need thereof.
 6. Apharmaceutical composition, comprising: the compound of claim 1, and apharmaceutically acceptable carrier.
 7. The method of claim 5, whereinthe estrogen receptor related treatment is contraception.
 8. The methodof claim 5, wherein the estrogen receptor related treatment is treatmentof benign prostate hypertrophy.
 9. The method of claim 5, wherein theestrogen receptor related treatment is treatment of osteoporosis. 10.The method of claim 5, wherein the estrogen receptor related treatmentis treatment of Alzheimer's disease.
 11. The method of claim 5, whereinthe estrogen receptor related treatment is treatment of depression.